乳腔镜前哨淋巴结活检及腋窝淋巴结清扫的临床研究

目的：探讨乳腺癌在乳腔镜下行前哨淋巴结活检及腋窝淋巴结清扫的可行性。方法：通过亚甲蓝示踪对40例Ⅰ、Ⅱ期乳腺癌行乳腔镜前哨淋巴结活检（ESLNB）,然后行乳腔镜腋窝淋巴结清扫（EALND）,对获得的全部淋巴结行病理检查HE染色,确定前哨淋巴结（SLN）检出率、假阴性率等。结果：40例乳腺癌患者SLN检出率为97.44%（39/40）,准确率为94.87%（37/39）,灵敏度为94.74%（18/19）,假阴性率5.26%（1/19）;每例平均前哨淋巴结活检（SLNB）检出数目1-6枚,腋窝淋巴结清扫（ALND）检出数目10-29枚。结论：应用乳腔镜下前哨淋巴结活检和腋窝淋巴结清扫准确可行,美容效果好,并发症低,可对早期乳腺癌进行准确腋窝淋巴结分期。     

酸性染料比色法测定牛大力中总生物碱的含量

目的 建立牛大力总生物碱的测定方法,测定牛大力总生物碱的含量.方法 采用酸性染料比色法测定,并对线性范围、加样回收率、精密度、专属性、稳定性等项目进行了方法学考察.结果 测定方法线性范围8.1～121.5 μg,相关系数为0.9998,平均加样回收率为98.2％,具有良好的精密度.结论 该含量测定方法操作简单,灵敏度高,准确性好,重现性好,可用于测定牛大力中总生物碱的含量.     

ED2000®: 585 nm collagen remodelling pulsed dye laser

The wavelength of 585nm corresponds to an absorption peak of haemoglobin. The heating effect in these skin layers triggers the release of various growth factors that stimulate collagen remodelling and tightening. We report our experience with a 585nm collagen remodelling, double flashlamp excited pumped dye laser was used (ED2000^®, Deka MELA, Calenzano, Italy), spot size 5?mm, energy density (fluence J/cm²) from 2 to 4?J/cm², emission modality (repetition rate) at 0.5?Hz, with a short pulse duration of 250?μsec. The efficiency of 585?nm collagen remodelling pulsed dye laser is controversial in only one session. It is probably reasonable to inform patients that 3–4 treatment sessions are necessary, and that 10% of the patient have no response to nonablative photorejuvenation.

Because of its low fluence and is shorter pulse duration, the 585?nm collagen remodelling pulsed dye laser has limited efficacy for the treatment of port wine stains. However, it may offer patients with erythematous, raised or hypertrophic acne scars or striae distensae a permanent cosmetic solution. This laser is safe and effective in the treatment of surgical scars starting as soon as possible, on the day of suture removal if possible. We found that 96.3% of molluscum contagiosum healed after the first treatment, the other 3.7% after the second.     

Evolving methods for single nucleotide polymorphism detection: Factor V Leiden mutation detection

Background: The many techniques used to diagnose the Factor V Leiden (FVL) mutation, the most common hereditary hypercoagulation disorder in Eurasians, and the most frequently requested genetic test reflect the evolving strategies in protein and DNA diagnosis. Methods: Here, molecular methods to diagnose the FVL mutation are discussed. Results: Protein‐based detection assays include the conventional functional activated protein C resistance coagulation test and the recently reported antibody‐mediated sensor detection; and DNA‐based assays include approaches that use electrophoretic fractionation e.g., restriction fragment length polymorphism, denaturing gradient gel electrophoresis, and single‐stranded conformational PCR analysis, DNA hybridization (e.g., microarrays), DNA polymerase‐based assays, e.g., extension reactions, fluorescence polarization template‐directed dye‐terminator incorporation, PCR assays (e.g., amplification‐refractory mutation system, melting curve analysis using real‐time quantitative PCR, and helicase‐dependent amplification), DNA sequencing (e.g., direct sequencing, pyrosequencing), cleavase‐based Invader assay and ligase‐based assays (e.g., oligonucleotide ligation assay and ligase‐mediated rolling circle amplification). Conclusion: The method chosen by a laboratory to diagnose FVL not only depends on the available technical expertise and equipment, but also the type, variety, and extent of other genetic disorders being diagnosed. J. Clin. Lab. Anal. 25:259–288, 2011. © 2011 Wiley‐Liss, Inc.     

混合均匀试验优选飞龙掌血散剂总生物碱提取工艺

目的:优选飞龙掌血散剂中总生物碱的提取工艺,并建立总生物碱含量测定方法。方法:以加水量、提取时间和提取次数为考察因素,以总生物碱含量为评价指标,采用混合均匀试验优选提取工艺,并采用酸性染料比色法测定总生物碱含量。结果:最佳提取工艺为加入12倍量水,提取2次,每次90min,该工艺下总生物碱含量为1.53%。结论:所选工艺和含量测定方法合理、可行,可为飞龙掌血的进一步研究提供参考。     

Arsenite-induced autophagy is associated with proteotoxicity in human lymphoblastoid cells

Epidemiological studies of arsenic-exposed populations have provided evidence that arsenic exposure in humans is associated with immunosuppression. Previously, we have reported that arsenite-induced toxicity is associated with the induction of autophagy in human lymphoblastoid cell lines (LCL). Autophagy is a cellular process that functions in the degradation of damaged cellular components, including protein aggregates formed by misfolded or damaged proteins. Accumulation of misfolded or damaged proteins in the endoplasmic reticulum (ER) lumen causes ER stress and activates the unfolded protein response (UPR). In an effort to investigate the mechanism of autophagy induction by arsenite in the LCL model, we examined the potential contribution of ER stress and activation of the UPR. LCL exposed to sodium arsenite for 8-days induced expression of UPR-activated genes, including CHOP and GRP78, at the RNA and the protein level. Evidence for activation of the three arms of the UPR was observed. The arsenite-induced activation of the UPR was associated with an accumulation of protein aggregates containing p62 and LC3, proteins with established roles in the sequestration and autophagic clearance of protein aggregates. Taken together, these data provide evidence that arsenite-induced autophagy is associated with the generation of ER stress, activation of the UPR, and formation of protein aggregates that may be targeted to the lysosome for degradation.     

厌氧污泥法处理偶氮染料废水的初步研究

目的:利用驯化的强化厌氧污泥,对模拟偶氮染料废水进行了厌氧生物处理研究。结果:厌氧强化污泥对甲基橙能进行快速、高效、彻底的降解,对甲基橙的浓度耐受也很高,1000mg/L的甲基橙144h内能降解98%;对甲基橙、甲基红、刚果红、橙黄Ⅰ和铬黑T连续投加40d依然能达到基本降解的目的(80%以上),具有广谱性和降解稳定性。     

Synthesis and in vitro cytotoxic evaluation of novel diazaspiro bicyclo hydantoin derivatives in human leukemia cells: A SAR study

Summary  To study the structure activity relationship (SAR) on the cytotoxic activity and probe the structural requirement for the potent antitumor activity, a series of novel diazaspiro bicyclo hydantoin derivatives were designed and synthesized. Their structures were confirmed by ¹H NMR, LCMS and IR analyses. The antiproliferative effect of these compounds were determined against human leukemia, K562 (chronic myelogenous leukemia) and CEM (T-cell leukemia) cells using trypan blue and MTT assay, and the SAR associated with the position of N-terminal substituents in diazaspiro bicyclo hydantoin have also been discussed. It has been observed that these compounds displayed strong, moderate and weak cytotoxic activities. Interestingly, compounds having electron withdrawing groups at third and fourth position of the phenyl ring displayed selectively cytotoxic activities to both the cell lines tested with IC₅₀ value lower than 50 μM. In addition, the cytotoxic activities of the compounds 7(a–o) bearing the substituents at N−3 position of diazaspiro bicyclo hydantoin increases in the order alkene > ester > ether and plays an important role in determining their antitumor activities. The position and number of substituents in benzyl group attached to N−8 of diazaspiro bicyclo hydantoin nucleus interacted selectively with specific targets leading to the difference of biochemical and pharmacological effects.     

Oxidative stress induction by T-2 toxin causes DNA damage and triggers apoptosis via caspase pathway in human cervical cancer cells

T-2 toxin is the most toxic trichothecene and both humans and animals suffer from several pathological conditions after consumption of foodstuffs contaminated with trichothecenes. We investigated the molecular mechanism of T-2 toxin induced cytotoxicity and cell death in HeLa cells. T-2 toxin at LC50 of 10 ng/ml caused time dependent increase in cytotoxicity as assessed by dye uptake, lactatedehydrogenase leakage and MTT assay. The toxin caused generation of reactive oxygen species as early as 30 min followed by significant depletion of glutathione levels and increased lipid peroxidation. The results indicate oxidative stress as underlying mechanism of cytotoxicity. Single stranded DNA damage after T-2 treatment was observed as early as 2 and 4 h by DNA diffusion assay. The cells exhibited apoptotic morphology like condensed chromatin and nuclear fragmentation after 4 h of treatment. Downstream of T-2 induced oxidative stress and DNA damage a time dependent increase in expression level of p53 protein was observed. The increase in Bax/Bcl2 ratio indicated shift in response, in favour of apoptotic process in T-2 toxin treated cells. Western blot analysis showed increase in levels of mitochondrial apoptogenic factors Bax, Bcl-2, cytochrome-c followed by activation of caspases-9, -3 and -7 leading to DNA fragmentation and apoptosis. In addition to caspase-dependent pathway, our results showed involvement of caspase-independent AIF pathway in T-2 induced apoptosis. Broad spectrum caspase inhibitor z-VAD-fmk could partially protect the cells from DNA damage but could not inhibit AIF induced oligonucleosomal DNA fragmentation beyond 4 h. Results of the study clearly show that oxidative stress is the underlying mechanism by which T-2 toxin causes DNA damage and apoptosis.